Compounds for treating disorders where a decreased level of plasma FFA is desired

ABSTRACT

The present invention relates to novel compounds, compositions containing them, and their use for treating medical disorders where a decreased level of plasma free fatty acids (FFA) is desired.

FIELD OF INVENTION

[0001] The present invention relates to novel compounds, compositionscontaining them, and their use for treating medical disorders where adecreased level of plasma free fatty acids (FFA) is desired.

BACKGROUND OF THE INVENTION

[0002] The overall energy homeostasis of a mammalian system requires ahigh degree of regulation to ensure the availability of the appropriatesubstrate at the appropriate time. Plasma glucose levels rise during thepost-prandial state, to return to pre-prandial levels within 2-3 hours.During these 2-3 hours, insulin promotes glucose uptake by skeletalmuscle and adipose tissue and decreases the release of free fatty acids(FFA) from adipocytes, to ensure that the two substrates do not competewith each other. When plasma glucose levels fall, an elevation in plasmaFFA is necessary to switch from glucose to fat utilization by thevarious tissues.

[0003] In individuals with insulin resistance, FFA levels do not fall inresponse to insulin, as they do in normal individuals, preventing thenormal utilization of glucose by skeletal muscle, adipose and liver.Furthermore, there is a negative correlation between insulin sensitivityand plasma FFA levels.

[0004] Hormone-sensitive lipase (HSL) is an enzyme, expressed primarilyin adipocytes, that catalyses the conversion of triglycerides toglycerol and fatty acids. It is through the regulation of this enzymethat the levels of circulating FFA are modulated. Insulin leads to theinactivation of HSL with a subsequent fall in plasma FFA levels duringthe post-prandial state, followed by the activation of the enzyme whenthe insulin concentration falls and catecholamines rise during thepost-absorptive period. The activation of HSL leads to an increase inplasma FFA, as they become the main source of energy during fasting.

[0005] The activation-inactivation of HSL is mediated through thecAMP-protein kinase A and AMP-dependent kinase pathways. There arecompounds, such as nicotinic acid and its derivatives, that decrease theactivation of HSL via these pathways and cause a decrease in lipolysisthat leads to a reduction in the FFA levels. These drugs have abeneficial effect in the utilization of glucose and in the normalizationof the excess triglyceride synthesis seen in patients with elevated FFA.However, since these pathways are used by other processes in the body,these drugs have severe side effects.

[0006] We have now found compounds that specifically inhibit thelipolytic activity of HSL and lead to a decrease in plasma FFA levels.These compounds can be used to treat disorders where a decreased levelof plasma FFA is desired, such as insulin resistance, dyslipidemia andabnormalities of lipoprotein metabolism.

[0007] One object of the present invention is thus to provide compoundsthat specifically inhibit the lipolytic activity of HSL. A furtherobject is to provide compounds which have good oral bioavailability.

SUMMARY OF THE INVENTION

[0008] One aspect of the present invention thus provides novel compoundsthat specifically inhibit the lipolytic activity of HSL.

[0009] These compounds can be utilized in vitro as unique research toolsfor understanding, inter alia, how the lipolytic activity of HSL isregulated at the adipocyte level.

[0010] Moreover, the compounds can also be administered in vivo forinhibiting the lipolytic activity of HSL.

DESCRIPTION OF THE INVENTION

[0011] Accordingly, the present invention relates to a compound ofgeneral formula I

[0012] wherein

[0013] A is a nitrogen containing ring system attached through thenitrogen atom, which nitrogen containing ring system is optionallysubstituted with one or more substituents independently selected fromhalogen, hydroxy, amino, oxy, cyano, nitro, C₁₋₆-alkyl and C₁₋₆-alkoxy;

[0014] wherein each of the C₁₋₆-alkyl or C₁₋₆-alkoxy may optionally besubstituted with one or more substituents independently selected fromhydroxy, halogen, amino, cyano and nitro;

[0015] R¹ is hydrogen or C₁₋₆-alkyl;

[0016] X¹ is —S— or —O—;

[0017] X² is —S— or —O—;

[0018] B is aryl optionally substituted with one or more substituentsindependently selected from halogen, hydroxy, amino, cyano, nitro,C₁₋₆-alkyl, C₁₋₆-alkoxy, C₁₋₆-alkylcarbonyl, C₁₋₆-alkoxycarbonyl,C₃₋₁₀-cycloalkyl, aryl, aryl-C₁₋₆-alkyl, aryl-C₁₋₆-alkoxy,aryl-C₁₋₆-alkoxycarbonyl, arylcarbonyl, —NR²⁰—C(═O)—C₁₋₆-alkyl,—NR²⁰—C(═O)—C₁₋₆-alkoxy, —NR²⁰—C₁₋₆-alkyl and—C₁₋₆-alkyl-NR²⁰—C₁₋₆-alkyl;

[0019] wherein each of the C₁₋₆-alkyl, C₁₋₆-alkoxy, C₃₋₁₀-cycloalkyl oraryl may optionally be substituted with one or more substituentsindependently selected from hydroxy, halogen, amino, cyano and nitro;

[0020] R²⁰ is hydrogen or C₁₋₆-alkyl;

[0021] or B is

[0022] wherein

[0023] Y¹ is —C(R²¹)═ or —N═;

[0024] Y² is —C(R²²)═ or —N═;

[0025] Y³ is —C(R²³)═ or —N═;

[0026] Y⁴ is —C(R²⁴)═ or —N═;

[0027] wherein R²¹, R²², R²³ and R²⁴ independently are hydrogen,halogen, hydroxy, amino, cyano, nitro, C₁₋₆-alkyl, C₁₋₆-alkoxy,C₁₋₆-alkylcarbonyl, C₁₋₆-alkoxycarbonyl, C₃₋₁₀-cycloalkyl, aryl,aryl-C₁₋₆-alkyl, aryl-C₁₋₆-alkoxy, aryl-C₁₋₆-alkoxycarbonyl,arylcarbonyl, —NR²⁵—C(═O)—C₁₋₆-alkyl, —NR²⁵—C(═O)—C₁₋₆-alkoxy,—NR²⁵—C₁₋₆-alkyl or —C₁₋₆-alkyl-NR²⁵—C₁₋₆-alkyl;

[0028] wherein each of the C₁₋₆-alkyl, C₁₋₆-alkoxy, C₃₋₁₀-cycloalkyl, oraryl may optionally be substituted with one or more substituentsindependently selected from hydroxy, halogen, amino, cyano and nitro;

[0029] R²⁵ is hydrogen or C₁₋₆-alkyl;

[0030] or a pharmaceutically acceptable salt thereof.

[0031] Moreover, the compounds of formula I may comprise any opticalisomers thereof, in the form of separated, pure or partially purifiedoptical isomers or racemic mixtures thereof.

[0032] Whenever one or more chiral carbon atoms are present, such chiralcenter or centers may be in the R- or S-configuration, or a mixture of Rand S.

[0033] In one embodiment of the compound of formula I, A is

[0034] wherein

[0035] R⁷, R⁸, and R⁹ independently are hydrogen or C₁₋₆-alkyl;

[0036] R¹⁰, R¹¹, R¹², and R¹³ independently are hydrogen, halogen,C₁₋₆-alkyl or C₁₋₆-alkoxy.

[0037] In one embodiment R⁷ is hydrogen. In a second embodiment R⁷ isC₁₋₆-alkyl, in particular methyl. In a third embodiment R⁸ is hydrogen.In a further embodiment R⁸ is C₁₋₆-alkyl, in particular methyl. In astill further embodiment R⁹ is hydrogen. In a further embodiment R¹⁰ ishydrogen. In a still further embodiment R¹¹ is hydrogen. In a furtherembodiment R¹¹ is C₁₋₆-alkoxy, in particular methoxy. In a still furtherembodiment R¹² is hydrogen. In a further embodiment R¹² is C₁₋₆-alkoxy,in particular methoxy. In a still further embodiment R¹³ is hydrogen.

[0038] In a further embodiment of the compound of formula I, A is

[0039] wherein R¹⁴, R¹⁵, R¹⁶, R¹⁷ and R¹⁸ independently are hydrogen,halogen, C₁₋₆-alkyl or C₁₋₆-alkoxy.

[0040] In one embodiment A is

[0041] In a second embodiment A is

[0042] In a third embodiment A is

[0043] In a further embodiment A is

[0044] In a still further embodiment R¹⁴ is hydrogen. In a furtherembodiment R¹⁵ is hydrogen. In a still further embodiment R¹⁶ ishydrogen. In a further embodiment R¹⁷ is hydrogen. In a still furtherembodiment R¹⁸ is hydrogen.

[0045] In a still further embodiment of the compound of formula I, inthe nitrogen containing ring system A at least one of the atoms next tothe nitrogen atom through which A is attached is unsubstituted.

[0046] In the compound of the above formula I, A is preferably3,4-dihydro-1H-isoquinolin-2-yl,6,7-dimethoxy-3,4-dihydro-1H-isoquinolin-2-yl,1-methyl-3,4-dihydro-1H-isoquinolin-2-yl,3-methyl-3,4-dihydro-1H-isoquinolin-2-yl, piperidin-1-yl ormorpholin-4-yl.

[0047] In a still further embodiment of the compound of formula I, R¹ ishydrogen. In a further embodiment R¹ is C₁₋₆-alkyl, in particularmethyl.

[0048] In a still further embodiment of the compound of formula I, X¹ is—S—. In a further embodiment X¹ is —O—.

[0049] In a still further embodiment of the compound of formula I, X² is—S—. In a further embodiment X² is —O—.

[0050] In a still further embodiment of the compound of formula I, X¹ is—O— and X² is —O—. In a further embodiment of the compound of formula I,X¹ is —S— and X² is —S—.

[0051] In a still further embodiment of the compound of formula I, B isaryl optionally substituted with one or more substituents independentlyselected from

[0052] halogen, hydroxy, amino, cyano, nitro, C₁₋₆-alkyl, C₁₋₆-alkoxy,C₁₋₆-alkylcarbonyl, C₁₋₆-alkoxycarbonyl, C₃₋₁₀-cycloalkyl, aryl,aryl-C₁₋₆-alkyl, aryl-C₁₋₆-alkoxy, aryl-C₁₋₆-alkoxycarbonyl,arylcarbonyl, —NR²⁰—C(═O)—C₁₋₆-alkyl, —NR²⁰—C(═O)—C₁₋₆-alkoxy,—NR²⁰—C₁₋₆-alkyl and —C₁₋₆-alkyl-NR²⁰—C₁₋₆-alkyl;

[0053] wherein each of the C₁₋₆-alkyl, C₁₋₆-alkoxy, C₃₋₁₀-cycloalkyl oraryl may optionally be substituted with one or more substituentsindependently selected from hydroxy, halogen, amino, cyano and nitro;

[0054] R²⁰ is hydrogen or C₁₋₆-alkyl.

[0055] In one embodiment, B is aryl. In a second embodiment, B is arylsubstituted with one or more halogen, in particular Br, F or Cl. In athird embodiment, B is aryl substituted with amino. In a furtherembodiment, B is aryl substituted with C₁₋₆-alkyl, such as tert-butyl,butyl-2-yl, 2-methyl-butyl-2-yl, propyl, ethyl or methyl. In a stillfurther embodiment, B is aryl substituted with C₁₋₆-alkoxy, such aspentoxy, butoxy, propoxy or methoxy, In a further embodiment, B is arylsubstituted with C₁₋₆-alkylcarbonyl, such as ethylcarbonyl ormethylcarbonyl. In a still further embodiment, B is aryl substitutedwith C₁₋₆-alkoxycarbonyl, such as ethoxycarbonyl or methoxycarbonyl. Ina further embodiment, B is aryl substituted with C₃₋₁₀-cycloalkyl, suchas cyclohexyl. In a still further embodiment, B is aryl substituted witharyl, such as phenyl. In a further embodiment, B is aryl substitutedwith aryl-C₁₋₆-alkyl, such as phenylmethyl. In a still furtherembodiment, B is aryl substituted with aryl-C₁₋₆-alkoxy, such asphenylmethoxy. In a further embodiment, B is aryl substituted witharyl-C₁₋₆-alkoxycarbonyl, such as phenylmethoxycarbonyl. In a stillfurther embodiment, B is aryl substituted with arylcarbonyl, such as4-methyl-phenylcarbonyl. In a further embodiment, B is aryl substitutedwith —NR²⁰—C(═O)—C₁₋₆-alkyl, such as methylcarbonylamino. In a stillfurther embodiment, B is aryl substituted with —NR²⁰—C(═O)—C₁₋₆-alkoxy,such as tert-butoxycarbonylamino. In a further embodiment, B is arylsubstituted with —NR²⁰—C₁₋₆-alkyl. In a still further embodiment, B isaryl substituted with —C₁₋₆-alkyl-NR²⁰—C₁₋₆-alkyl, such asdimethylaminoethyl.

[0056] In a still further embodiment of the compound of formula I, B is

[0057] wherein

[0058] Y¹ is —C(R²¹)═ or —N═;

[0059] Y² is —C(R²²)═ or —N═;

[0060] Y³ is —C(R²³)═ or —N═;

[0061] Y⁴ is —C(R²⁴)═ or —N═;

[0062] wherein R²¹, R²², R²³, and R²⁴ independently are hydrogen,halogen, hydroxy, amino, cyano, nitro, C₁₋₆-alkyl, C₁₋₆-alkoxy,C₁₋₆-alkylcarbonyl, C₁₋₆-alkoxycarbonyl, C₃₋₁₀-cycloalkyl,aryl-C₁₋₆-alkyl, aryl-C₁₋₆-alkoxy, aryl-C₁₋₆-alkoxycarbonyl,arylcarbonyl, —NR²⁵—C(═O)—C₁₋₆-alkyl, —NR²⁵—C(═O)—C₁₋₆-alkoxy,—NR²⁵—C₁₋₆-alkyl or —C₁₋₆-alkyl-NR²⁵—C₁₋₆-alkyl;

[0063] wherein each of the C₁₋₆-alkyl, C₁₋₆-alkoxy, C₃₋₁₀-cycloalkyl oraryl may optionally be substituted with one or more substituentsindependently selected from hydroxy, halogen, amino, cyano and nitro;

[0064] R²⁵ is hydrogen or C₁₋₆-alkyl.

[0065] In one embodiment, yt is —CH═. In a second embodiment, Y² is —N═.In a third embodiment, Y³ is —CH═. In a further embodiment Y⁴ is —N═.

[0066] In a still further embodiment of the compound of formula I, B is

[0067] wherein

[0068] R², R³, R⁴, R⁵ and R⁶ independently of each other are hydrogen,halogen, hydroxy, amino, cyano, nitro, C₁₋₆-alkyl, C₁₋₆-alkoxy,C₁₋₆-alkylcarbonyl, C₁₋₆-alkoxycarbonyl, C₃₋₁₀-cycloalkyl,aryl-C₁₋₆-alkyl, aryl-C₁₋₆-alkoxy, aryl-C₁₋₆-alkoxycarbonyl,arylcarbonyl, —NR²⁰—C(═O)—C₁₋₆-alkyl, —NR²⁰—C(═O)—C₁₋₆-alkoxy,—NR²⁰—C₁₋₆-alkyl or —C₁₋₆-alkyl-NR²⁰—C₁₋₆-alkyl;

[0069] wherein each of the C₁₋₆-alkyl, C₁₋₆-alkoxy, C₃₋₁₀-cycloalkyl oraryl may optionally be substituted with one or more substituentsindependently selected from hydroxy, halogen, amino, cyano and nitro;

[0070] R²⁰ is hydrogen or C₁₋₆-alkyl.

[0071] In a still further embodiment of the compound of formula I, B is

[0072] wherein R², R³, R⁴, R⁵ and R⁶ are defined as above. In oneembodiment, B is

[0073] such as 2-methyl-phenyl, 2-methoxy-phenyl, 2-bromo-phenyl,2-fluoro-phenyl. In a second embodiment, B is

[0074] such as 3-methyl-phenyl. In a further embodiment, B is

[0075] such as 4-methoxy-phenyl, 4-methoxy-carbonyl-phenyl,4-bromo-phenyl, 4-chloro-phenyl, 4-fluoro-phenyl, 4-methyl-phenyl,4-trifluoromethyl-phenyl, 4-tert-butoxycarbonylamino-phenyl,4-amino-phenyl, 4-phenyl-ethoxycarbonyl-phenyl, 4-propyl-phenyl,4-methoxy-amino-phenyl, 4-(4-methyl-carbonyl)-phenyl,4-phenyl-methyl-phenyl, 4-ethyl-phenyl, 4-phenyl-methoxy-phenyl,4-butoxy-phenyl, 4-(2-methyl-propyl-2-yl), 4-(propyl-2-yl),4-tert-butyl-phenyl, 4-dimethyl-amino-ethyl-phenyl,4-ethoxy-carbonyl-phenyl, 4-bi-phenyl, 4-pentoxy-phenyl,4-phenoxy-phenyl, 4-propoxy-phenyl, 4-triflouromethoxy-phenyl,4-phenyl-carbonyl-phenyl, 4-methoxy-carbonyl-methyl-phenyl,4-cyclohexyl-phenyl, 4-chloro-propyl-carbonyl-phenyl. In a still furtherembodiment, B is

[0076] In a further embodiment, B is

[0077] such as 2,4-dichloro-phenyl. In a still further embodiment, B is

[0078] In a further embodiment B is

[0079] In a still further embodiment, B is

[0080] In a further embodiment, B is

[0081] In a still further embodiment of the compound of formula I, B is

[0082] wherein R¹⁹ is hydrogen, C₁₋₆-alkyl, C₁₋₆-alkylcarbonyl,C₁₋₆-alkoxycarbonyl, aryl-C₁₋₆-alkyl, aryl-C₁₋₆-alkoxycarbonyl orarylcarbonyl. In one embodiment, B is 4-amino-phenyl. In a secondembodiment, B is 4-methyl-carbonyl-amino-phenyl.

[0083] Any possible combination of two or more of the embodimentsdescribed herein is comprised within the scope of the invention.

[0084] Preferred compounds of formula I of the invention are:

[0085] and pharmaceutically acceptable salts thereof.

[0086] The present invention also encompasses pharmaceuticallyacceptable salts of the present compounds. Such salts includepharmaceutically acceptable acid addition salts, pharmaceuticallyacceptable metal salts, ammonium and alkylated ammonium salts. Acidaddition salts include salts of inorganic acids as well as organicacids. Representative examples of suitable inorganic acids includehydrochloric, hydrobromic, hydroiodic, phosphoric, sulfuric, nitricacids and the like. Representative examples of suitable organic acidsinclude formic, acetic, trichloroacetic, trifluoroacetic, propionic,benzoic, cinnamic, citric, fumaric, glycolic, lactic, maleic, malic,malonic, mandelic, oxalic, picric, pyruvic, salicylic, succinic,methanesulfonic, ethanesulfonic, tartaric, ascorbic, pamoic,bismethylene salicylic, ethanedisulfonic, gluconic, citraconic,aspartic, stearic, palmitic, EDTA, glycolic, p-aminobenzoic, glutamic,benzenesulfonic, p-toluenesulfonic acids and the like. Further examplesof pharmaceutically acceptable inorganic or organic acid addition saltsinclude the pharmaceutically acceptable salts listed in J. Pharm. Sci.1977, 66, 2, which is incorporated herein by reference. Examples ofmetal salts include lithium, sodium, potassium, magnesium salts and thelike. Examples of ammonium and alkylated ammonium salts includeammonium, methylammonium, dimethylammonium, trimethylammonium,ethylammonium, hydroxyethylammonium, diethylammonium, butylammonium,tetramethylammonium salts and the like.

[0087] Also intended as pharmaceutically acceptable acid addition saltsare the hydrates which the present compounds are able to form.

[0088] Any possible combination of two or more of the embodimentsdescribed herein is comprised within the scope of the present invention.

[0089] Pharmaceutical Composition

[0090] In a still further aspect, the invention relates to apharmaceutical composition comprising, as an active ingredient, acompound of general formula I

[0091] wherein

[0092] A is a nitrogen containing ring system attached through thenitrogen atom, which nitrogen containing ring system is optionallysubstituted with one or more substituents independently selected fromhalogen, hydroxy, amino, oxy, cyano, nitro, C₁₋₆-alkyl and C₁₋₆-alkoxy;

[0093] wherein each of the C₁₋₆-alkyl, or C₁₋₆-alkoxy may optionally besubstituted with one or more substituents independently selected fromhydroxy, halogen, amino, cyano and nitro;

[0094] R¹ is hydrogen or C₁₋₆-alkyl;

[0095] X¹ is —S— or —O—;

[0096] X² is —S— or —O—;

[0097] B is aryl optionally substituted with one or more substituentsindependently selected from halogen, hydroxy, amino, cyano, nitro,C₁₋₆-alkyl, C₁₋₆-alkoxy, C₁₋₆-alkylcarbonyl, C₁₋₆-alkoxycarbonyl,C₃₋₁₀-cycloalkyl, aryl, aryl-C₁₋₆-alkyl, aryl-C₁₋₆-alkoxy,aryl-C₁₋₆-alkoxycarbonyl, arylcarbonyl, —NR²⁰—C(═O)—C₁₋₆-alkyl,—NR²⁰—C(═O)—C₁₋₆-alkoxy, —NR²⁰—C₁₋₆-alkyl and—C₁₋₆-alkyl—NR²⁰—C₁₋₆-alkyl;

[0098] wherein each of the C₁₋₆-alkyl, C₁₋₆-alkoxy, C₃₋₁₀-cycloalkyl oraryl may optionally be substituted with one or more substituentsindependently selected from hydroxy, halogen, amino, cyano and nitro;

[0099] R²⁰ is hydrogen or C₁₋₆-alkyl;

[0100] or B is

[0101] wherein

[0102] Y¹ is —C(R²¹)═ or —N═;

[0103] Y² is —C(R²²)═ or —N═;

[0104] Y³ is —C(R²³)═ or —N═;

[0105] Y⁴ is —C(R²⁴)═ or —N═;

[0106] wherein R²¹, R²², R²³ and R²⁴ independently are hydrogen,halogen, hydroxy, amino, cyano, nitro, C₁₋₆-alkyl, C₁₋₆-alkoxy,C₁₋₆-alkylcarbonyl, C₁₋₆-alkoxycarbonyl, C₃₋₁₀-cycloalkyl, aryl,aryl-C₁₋₆-alkyl, aryl-C₁₋₆-alkoxy, aryl-C₁₋₆-alkoxycarbonyl,arylcarbonyl, —NR²⁵—C(═O)—C₁₋₆-alkyl, —NR²⁵—C(═O)—C₁₋₆-alkoxy,—NR²⁵—C₁₋₆-alkyl or —C₁₋₆-alkyl-NR²⁵—C₁₋₆-alkyl;

[0107] wherein each of the C₁₋₆-alkyl, C₁₋₆-alkoxy, C₃₋₁₀-cycloalkyl oraryl may optionally be substituted with one or more substituentsindependently selected from hydroxy, halogen, amino, cyano and nitro;

[0108] R²⁵ is hydrogen or C₁₋₆-alkyl; or a pharmaceutically acceptablesalt thereof together with a pharmaceutically acceptable carrier.

[0109] The compounds of the invention may be administered alone or incombination with pharmaceutically acceptable carriers or excipients, ineither single or multiple doses. The pharmaceutical compositionsaccording to the invention may be formulated with pharmaceuticallyacceptable carriers or diluents as well as any other known adjuvants andexcipients in accordance with conventional techniques such as thosedisclosed in Remington: The Science and Practice of Pharmacy, 19^(th)Edition, Gennaro, Ed., Mack Publishing Co., Easton, Pa., 1995.

[0110] The pharmaceutical compositions may be specifically formulatedfor administration by any suitable route such as the oral, rectal,nasal, pulmonary, topical (including buccal and sublingual),transdermal, intracisternal, intraperitoneal, vaginal and parenteral(including subcutaneous, intramuscular, intrathecal, intravenous andintradermal) route, the oral route being preferred. It will beappreciated that the preferred route will depend on the generalcondition and age of the subject to be treated, the nature of thecondition to be treated and the active ingredient chosen.

[0111] Pharmaceutical compositions for oral administration include soliddosage forms such as capsules, tablets, dragees, pills, lozenges,powders and granules. Where appropriate, they can be prepared withcoatings such as enteric coatings or they can be formulated so as toprovide controlled release of the active ingredient such as sustained orprolonged release according to methods well known in the art.

[0112] Liquid dosage forms for oral administration include solutions,emulsions, suspensions, syrups and elixirs.

[0113] Pharmaceutical compositions for parenteral administration includesterile aqueous and non-aqueous injectable solutions, dispersions,suspensions or emulsions as well as sterile powders to be reconstitutedin sterile injectable solutions or dispersions prior to use. Depotinjectable formulations are also contemplated as being within the scopeof the present invention.

[0114] Other suitable administration forms include suppositories,sprays, ointments, cremes, gels, inhalants, dermal patches, implants,etc.

[0115] The therapeutic dose of the compound will depend upon thefrequency and mode of administration, the sex, age, weight and generalcondition of the subject treated, the nature and severity of thecondition treated and any concomitant diseases to be treated, and otherfactors evident to those skilled in the art. The formulations mayconveniently be presented in unit dosage form by methods known to thoseskilled in the art. In one embodiment, the composition in unit dosageform comprises from about 0.05 to about 2000 mg, preferably from about0.1 to about 500 mg of the compound of formula I or a pharmaceuticallyacceptable salt thereof.

[0116] In a still further embodiment, the pharmaceutical composition isfor oral, nasal, transdermal, pulmonary or parenteral administration.

[0117] For parenteral routes, such as intravenous, intrathecal,intramuscular and similar administration, typical doses are of the orderof about half the dose employed for oral administration.

[0118] The compounds of this invention are generally utilized as thefree substance or as a pharmaceutically acceptable salt thereof. Oneexample is an acid addition salt of a compound having the a free basefunctionality. When a compound of the invention contains a free basefunctionality, such salts are prepared in a conventional manner bytreating a solution or suspension of the free base form of the compoundwith a chemical equivalent of a pharmaceutically acceptable acid, forexample an inorganic or organic acid. Representative examples hereof arementioned above. Physiologically acceptable salts of a compound with ahydroxy group include the anion of said compound in combination with asuitable cation, such as sodium or ammonium ion.

[0119] For parenteral administration, solutions of the present compoundsin sterile aqueous solution, aqueous propylene glycol or sesame orpeanut oil may be employed. Such aqueous solutions should be suitablebuffered if necessary and the liquid diluent first rendered isotonicwith sufficient saline or glucose. The aqueous solutions areparticularly suitable for intravenous, intramuscular, subcutaneous andintraperitoneal administration. The sterile aqueous media employed areall readily available by standard techniques known to those skilled inthe art.

[0120] Suitable pharmaceutical carriers include inert solid diluents orfillers, sterile aqueous solution and various organic solvents. Examplesof solid carriers are lactose, terra alba, sucrose, cyclodextrin, talc,gelatine, agar, pectin, acacia, magnesium stearate, stearic acid orlower alkyl ethers of cellulose. Examples of liquid carriers are syrup,peanut oil, olive oil, phospholipids, fatty acids, fatty acid amines,polyoxyethylene or water. Similarly, the carrier or diluent may includeany sustained release material known in the art, such as glycerylmonostearate or glyceryl distearate, alone or mixed with a wax. Thepharmaceutical compositions formed by combining the compounds of theinvention and the pharmaceutically acceptable carriers are then readilyadministered in a variety of dosage forms suitable for the disclosedroutes of administration. The formulations may conveniently be presentedin unit dosage form by methods known in the art of pharmacy.

[0121] Formulations of the present invention suitable for oraladministration may be presented as discrete units such as capsules ortablets, each containing a predetermined amount of the activeingredient, and which may include a suitable excipient. Theseformulations may be in the form of powder or granules, as a solution orsuspension in an aqueous or non-aqueous liquid, or as an oil-in-water orwater-in-oil liquid emulsion.

[0122] If a solid carrier is used for oral administration, thepreparation may be tabletted, placed in a hard gelatine capsule inpowder or pellet form or it can be in the form of a troche or lozenge.The amount of solid carrier will vary widely but will usually be fromabout 25 mg to about 1 g. If a liquid carrier is used, the preparationmay be in the form of a syrup, emulsion, soft gelatine capsule orsterile injectable liquid such as an aqueous or non-aqueous liquidsuspension or solution.

[0123] A typical tablet which may be prepared by conventional tablettingtechniques may contain: Core: Active compound (as free compound or saltthereof)   5 mg Colloidal silicon dioxide (Aerosil ™) 1.5 mg Cellulose,microcryst. (Avicel ™)  70 mg Modified cellulose gum (Ac-Di-Sol ™) 7.5mg Magnesium stearate q.s. Coating: Hydroxypropylmethylcellulose (HPMC)approx.   9 mg *Mywacett ™ 9-40 T approx. 0.9 mg

[0124] The compounds of the invention may be administered to a mammal,especially a human, in need thereof. Such mammals include also animals,both domestic animals, e.g. household pets, and non-domestic animalssuch as wildlife.

[0125] In a further aspect of the invention the present compounds may beadministered in combination with further pharmacologically activesubstances e.g. an antidiabetic or other pharmacologically activematerial, including other compounds for the treatment and/or preventionof insulin resistance and diseases, wherein insulin resistance is thepathophysiological mechanism.

[0126] Furthermore, the compounds according to the invention may beadministered in combination with antiobesity agents or appetiteregulating agents.

[0127] In a further embodiment the invention relates to a pharmaceuticalcomposition for specifically inhibiting the lipolytic activity of HSL,the composition comprising, as an active ingredient, a compound offormula I or a pharmaceutically acceptable salt thereof together with apharmaceutically acceptable carrier or diluent.

[0128] It has been demonstrated that compounds of the general formula Ipossess the ability to specifically inhibit HSL in vivo. The compoundsmay therefore be used in the treatment of conditions which requiredecreased plasma FFA.

[0129] In a still further aspect, the present invention relates to theuse of of a compound that specifically inhibits the lipolytic activityof HSL, or a pharmaceutically acceptable salt thereof, for thepreparation of a medicament for the treatment of a disorder where adecreased level of plasma FFA is desired.

[0130] In one embodiment, the disorder where a decreased level of plasmaFFA is desired is diabetes type 2, insulin resistance, impaired glucosetolerance, hyperglycemia, dyslipidemia, or abnormalities of lipoproteinmetabolism.

[0131] In a further embodiment, the compound that specifically inhibitslipolytic activity of HSL is a compound of general formula I

[0132] wherein

[0133] A is a nitrogen containing ring system attached through thenitrogen atom, which nitrogen containing ring system is optionallysubstituted with one or more substituents independently selected fromhalogen, hydroxy, amino, oxy, cyano, nitro, C₁₋₆-alkyl and C₁₋₆-alkoxy;

[0134] wherein each of the C₁₋₆-alkyl or C₁₋₆-alkoxy may optionally besubstituted with one or more substituents independently selected fromhydroxy, halogen, amino, cyano and nitro;

[0135] R¹ is hydrogen or C₁₋₆-alkyl;

[0136] X¹ is —S— or —O—;

[0137] X² is —S— or —O—;

[0138] B is aryl optionally substituted with one or more substituentsindependently selected from halogen, hydroxy, amino, cyano, nitro,C₁₋₆-alkyl, C₁₋₆-alkoxy, C₁₋₆-alkylcarbonyl, C₁₋₆-alkoxycarbonyl,C₃₋₁₀-cycloalkyl, aryl, aryl-C₁₋₆-alkyl, aryl-C₁₋₆-alkoxy,aryl-C₁₋₆-alkoxycarbonyl, arylcarbonyl, —NR²⁰—C(═O)—C₁₋₆-alkyl,—NR²⁰—C(═O)—C₁₋₆-alkoxy, —NR²⁰—C₁₋₆-alkyl and—C₁₋₆alkyl-NR²⁰—C₁₋₆-alkyl;

[0139] wherein each of the C₁₋₆-alkyl, C₁₋₆-alkoxy, C₃₋₁₀-cycloalkyl oraryl may optionally be substituted with one or more substituentsindependently selected from hydroxy, halogen, amino, cyano and nitro;

[0140] R²⁰ is hydrogen or C₁₋₆-alkyl;

[0141] or B is

[0142] wherein

[0143] Y¹ is —C(R²¹)═ or —N═;

[0144] Y² is —C(R²²)═ or —N═;

[0145] Y³ is —C(R²³)═ or —N═;

[0146] Y⁴ is —C(R²⁴)═ or —N═;

[0147] wherein R²¹, R²², R²³ and R²⁴ independently are hydrogen,halogen, hydroxy, amino, cyano, nitro, C₁₋₆-alkyl, C₁₋₆-alkoxy,C₁₋₆-alkylcarbonyl, C₁₋₆-alkoxycarbonyl, C₃₋₁₀-cycloalkyl, aryl,aryl-C₁₋₆alkyl, aryl-C₁₋₆-alkoxy, aryl-C₁₋₆-alkoxycarbonyl,arylcarbonyl, —NR²⁵—C(═O)—C₁₋₆-alkyl, —NR²⁵—C(═O)—C₁₋₆-alkoxy,—NR²⁵—C₁₋₆-alkyl or —C₁₋₆-alkyl-NR²⁵—C₁₋₆-alkyl;

[0148] wherein each of the C₁₋₆-alkyl, C₁₋₆-alkoxy, C₃₋₁₀-cycloalkyl oraryl may optionally be substituted with one or more substituentsindependently selected from hydroxy, halogen, amino, cyano and nitro;

[0149] R²⁵ is hydrogen or C₁₋₆-alkyl;

[0150] or a pharmaceutically acceptable salt thereof, together with apharmaceutically acceptable carrier.

[0151] In a still further aspect, the invention relates to a method oftreating disorders of a mammal, e.g. a human, where a decreased level ofplasma FFA is desired, the method comprising administering to saidmammal an effective amount of a compound that specifically inhibits thelipolytic activity of HSL, or a pharmaceutically acceptable saltthereof.

[0152] In one embodiment of the method, the disorder where a decreasedlevel of plasma FFA is desired is diabetes type 2, insulin resistance,impaired glucose tolerance, hyperglycemia, dyslipidemia, orabnormalities of lipoprotein metabolism.

[0153] In a further embodiment, the compound that specifically inhibitslipolytic activity of HSL is a compound of general formula I

[0154] wherein

[0155] A is a nitrogen containing ring system attached through thenitrogen atom, which nitrogen containing ring system is optionallysubstituted with one or more substituents independently selected fromhalogen, hydroxy, amino, oxy, cyano, nitro, C₁₋₆-alkyl and C₁₋₆-alkoxy;

[0156] wherein each of the C₁₋₆-alkyl or C₁₋₆-alkoxy may optionally besubstituted with one or more substituents independently selected fromhydroxy, halogen, amino, cyano and nitro;

[0157] R¹ is hydrogen or C₁₋₆-alkyl;

[0158] X¹ is —S— or —O—;

[0159] X² is —S— or —O—;

[0160] B is aryl optionally substituted with one or more substituentsindependently selected from halogen, hydroxy, amino, cyano, nitro,C₁₋₆-alkyl, C₁₋₆-alkoxy, C₁₋₆-alkylcarbonyl, C₁₋₆-alkoxycarbonyl,C₃₋₁₀-cycloalkyl, aryl, aryl-C₁₋₆-alkyl, aryl-C₁₋₆-alkoxy,aryl-C₁₋₆-alkoxycarbonyl, arylcarbonyl, —NR²⁰—C(═O)—C₁₋₆-alkyl,—NR²⁰—C(═O)—C₁₋₆-alkoxy, —NR²⁰—C₁₋₆-alkyl and—C₁₋₆-alkyl-NR²⁰—C₁₋₆-alkyl;

[0161] wherein each of the C₁₋₆-alkyl, C₁₋₆-alkoxy, C₃₋₁₀-cycloalkyl oraryl may optionally be substituted with one or more substituentsindependently selected from hydroxy, halogen, amino, cyano and nitro;

[0162] R²⁰ is hydrogen or C₁₋₆-alkyl;

[0163] or B is

[0164] wherein

[0165] Y¹ is —C(R²¹)═ or —N═;

[0166] Y² is —C(R²²)═ or —N═;

[0167] Y³ is —C(R²³)═ or —N═;

[0168] Y⁴ is —C(R²⁴)═ or —N═;

[0169] wherein R²¹, R²², R²³ and R²⁴ independently are hydrogen,halogen, hydroxy, amino, cyano, nitro, C₁₋₆alkyl, C₁₋₆-alkoxy,C₁₋₆-alkylcarbonyl, C₁₋₆-alkoxycarbonyl, C₃₋₁₀-cycloalkyl, aryl,aryl-C₁₋₆alkyl, aryl-C₁₋₆-alkoxy, aryl-C₁₋₆-alkoxycarbonyl,arylcarbonyl, —NR²⁵—C(═O)—C₁₋₆-alkyl, —NR²⁵—C(═O)—C₁₋₆-alkoxy,—NR²⁵—C₁₋₆-alkyl or —C₁₋₆-alkyl-NR²⁵—C₁₋₆-alkyl;

[0170] wherein each of the C₁₋₆-alkyl, C₁₋₆-alkoxy, C₃₋₁₀-cycloalkyl oraryl may optionally be substituted with one or more substituentsindependently selected from hydroxy, halogen, amino, cyano and nitro;

[0171] R²⁵ is hydrogen or C₁₋₆-alkyl;

[0172] or a pharmaceutically acceptable salt thereof, together with apharmaceutically acceptable carrier.

[0173] In a still further embodiment, the administration is carried outby the oral, nasal, transdermal, pulmonary or parenteral route.

[0174] In a still further aspect, the present invention relates to amethod of identifying compounds for the treatment of disorders where adecreased level of plasma FFA is desired, characterised by screening outcompounds that specifically inhibit HSL.

[0175] The specific inhibition of HSL can, e.g., be determined by theuse of the assays as described in the assays under “Pharmacologicalmethods” below.

[0176] In one embodiment of the invention, the compounds thatspecifically inhibit HSL give rise to at most about 20 percentinhibition, such as less than 20 percent inhibition, of the lipaseslipoprotein lipase (LPL) and hepatic lipase (HL) at a concentration of50 μM. In a further embodiment, the compounds that specifically inhibitHSL give rise to less than 10 percent inhibition of the lipases LPL andHL at a concentration of 50 μM.

[0177] In a still further embodiment, the compounds that specificallyinhibit HSL have an inhibition constant (with respect to inhibition ofHSL), K_(i), of less than 10 μM. In a further embodiment, the compoundsthat specifically inhibit HSL have a K_(i) (with respect to HSLinhibition) of less than 1 μM. K_(i) is defined in the normal manner,i.e.

K _(i)=([enzyme][inhibitor])/[enzyme-inhibitor complex]

[0178] where the terms in square brackets are equilibrium concentrationsof the species in question.

[0179] Within the context of the present invention, treatment is to beunderstood as treatment and/or prevention.

[0180] The present invention is further illustrated by the followingexamples; these examples are, however, not to be construed as limitingthe scope of protection.

[0181] Definitions

[0182] The following provides detailed definitions of the terms used todescribe the compounds of the invention:

[0183] “Halogen” designates an atom selected from the group consistingof F, Cl, Br and I.

[0184] The term “C₁₋₆-alkyl” in the present context designates asaturated, branched or straight hydrocarbon group having from 1 to 6carbon atoms. Representative examples include, but are not limited to,methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, sec-butyl,tert-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, n-hexyl,isohexyl and the like.

[0185] The term “C₁₋₆-alkoxy” in the present context designates a group—O—C₁₋₆-alkyl wherein C₁₋₆-alkyl is as defined above. Representativeexamples include, but are not limited to, methoxy, ethoxy, n-propoxy,isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, n-pentoxy,isopentoxy, neopentoxy, tert-pentoxy, n-hexoxy, isohexoxy and the like.

[0186] The term “C₃₋₁₀-cycloalkyl” as used herein represents a saturatedmono-, bi-, tri- or spiro-carbocyclic group having from 3 to 10 carbonatoms. Representative examples are cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl,bicyclo[3.2.1]octyl, spiro[4.5]decyl, norpinyl, norbonyl, norcaryl,adamantyl and the like.

[0187] The term “C₃₋₈-heterocyclyl” as used herein represents asaturated 3 to 8 membered ring containing one or more heteroatomsselected from nitrogen, oxygen and sulfur. Representative examples arepyrrolidyl, piperidyl, piperazinyl, morpholinyl, thiomorpholinyl,aziridinyl, tetrahydrofuranyl and the like.

[0188] The term “aryl” as used herein represents a carbocyclic aromaticring system being either monocyclic, bicyclic, or polycyclic, such asphenyl, biphenyl, naphthyl, anthracenyl, phenanthrenyl, fluorenyl,indenyl, pentalenyl, azulenyl, biphenylenyl and the like. Aryl is alsointended to include the partially hydrogenated derivatives of thecarbocyclic aromatic systems enumerated above. Non-limiting examples ofsuch partially hydrogenated derivatives are 1,2,3,4-tetrahydronaphthyl,1,4-dihydronaphthyl and the like.

[0189] The term “heteroaryl” as used herein represents a heterocyclicaromatic ring system containing one or more heteroatoms selected fromnitrogen, oxygen and sulfur such as furanyl, thiophenyl, pyrrolyl,oxazolyl, thiazolyl, imidazolyl, isoxazolyl, isothiazolyl,1,2,3-triazolyl, 1,2,4-triazolyl, pyranyl, pyridyl, pyridazinyl,pyrimidinyl, pyrazinyl, 1,2,3-triazinyl, 1,2,4-triazinyl,1,3,5-triazinyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl,1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-thiadiazolyl,1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, tetrazolyl,thiadiazinyl, indolyl, isoindolyl, benzofuranyl, benzothiophenyl(thianaphthenyl), indazolyl, benzimidazolyl, benzthiazolyl,benzisothiazolyl, benzoxazolyl, benzisoxazolyl, purinyl, quinazolinyl,quinolizinyl, quinolinyl, isoquinolinyl, quinoxalinyl, naphthyridinyl,pteridinyl, carbazolyl, azepinyl, diazepinyl, acridinyl and the like.Heteroaryl is also intended to include the partially hydrogenatedderivatives of the heterocyclic systems enumerated above. Non-limitingexamples of such partially hydrogenated derivatives are2,3-dihydrobenzofuranyl, 3,4-dihydroisoquinolinyl, pyrrolinyl,pyrazolinyl, indolinyl, oxazolidinyl, oxazolinyl, oxazepinyl and thelike.

[0190] The term “heterocyclic system” as used herein includes aromaticas well as non-aromatic ring moieties, which may be monocyclic, bicyclicor polycyclic, and containing in their ring structure one or moreheteroatoms selected from nitrogen, oxygen and sulfur. Non-limitingexamples of such heterocyclic systems are C₃₋₈-heterocyclyl andheteroaryl.

[0191] The term “nitrogen containing ring system” designates aheterocyclic system as defined herein having at least one nitrogen atom.

[0192] Certain of the above defined terms may occur more than once inthe structural formulae, and upon such occurrence each term shall bedefined independently of the other.

[0193] The term “optionally substituted” as used herein means that thegroups in question are either unsubstituted or substituted with one ormore of the substituents specified. When the groups in question aresubstituted with more than one substituent, the substituents may be thesame or different.

[0194] Pharmacological Methods

[0195] Compounds of formula I may be evaluated in vitro for theirefficacy and potency to specifically inhibit HSL, and such evaluationmay be performed as described below.

[0196] Assays

[0197] Hormone-sensitive lipase (HSL) [classified as a triacylglycerollipase, EC 3.1.1.3, in accordance with the Recommendations of theNomenclature Committee of the International Union of Biochemistry andMolecular Biology (available on the World Wide Web at the followingaddress: http://www.chem.qmw.ac.uk/iubmb/enzyme/)]

[0198] Materials. The Hormone-sensitive lipase employed was provided byDr. Cecilia Holm of Lund University, Sweden, or produced and purified byNovo Nordisk (NN) using the reagents and protocols employed by Dr. Holm.The substrates used are: ³H-labeled triolein (TO) from Amersham,Buckinghamshire, U.K. cat No. TRA191; 5-20 Ci/mmol dissolved in toluene,triolein (Sigma, Cat. No. T-1740) and mono-oleoyl-2-O-mono-oleylglycerol(MOME) synthesized at (NN) using the protocols provided by Dr. CeciliaHolm, Lund University, Sweden. Phosphatidyl choline (PC) andphosphatidyl inositol (PI) are from Sigma (St Luis Mo., cat. Nos. P-3556and P-5954, respectively). All other reagents are of commercial gradeand obtained from various commercial sources.

[0199] Methods. The preparation of the substrates is as follows:

[0200] TO: 30 μl PC:PI (20 mg/ml solution of PC:PI 3:1 prepared inchloroform)+128 μl cold TO+15 μl ³H-TO

[0201] MOME: 30 μl PC:PI+100 μl cold MOME+200 μl ³H-MOME

[0202] They are then evaporated under a gentle stream of N₂ followed by20-30 minutes in a Speedvac™ apparatus to ensure the absence of residualsolvent.

[0203] The samples are prepared as follows:

[0204] Blank: 100 μl PED-BSA

[0205] Samples/controls: 78 μl PED-BSA+20 μl inhibitor (variousconcentrations)+2 μl HSL (undiluted 0.12 mg/ml). If no inhibitors arebeing tested, the reaction contains 98 μl PED-BSA+2 μl HSL.

[0206] The samples are put on ice and then sonicated as follows:

[0207] With TO as substrate, add 2 ml potassium buffer (0.1M potassiumphosphate, pH 7) and then sonicate twice for 1 minute, with a 1-minuteinterval between cycles. Add an additional 1 ml potassium buffer andsonicate 4 times for 30 seconds, with a 30 second interval in betweencycles. To finish, add 1 ml BSA 20%-in potassium buffer, mix byinversion and store on ice until use.

[0208] With MOME as substrate: add 2 ml potassium buffer and thensonicate twice for 1 minute, with a 1 minute interval between cycles.Add 1.6 ml potassium buffer and sonicate 4 times for 30 seconds, with a30-second interval between cycles. To finish, add 0.4 ml BSA 20%-buffer,mix by inversion and store on ice until use.

[0209] The enzyme reaction is run at 37° C. The substrate mix is addedto the various reactions at 10 second intervals, incubated for 30minutes in a water bath and then stopped with 3.25 ml of a solutioncontaining CH₃OH:CHCl₃:heptane (10:9:7). After the addition of 1.05 mlpotassium buffer pH10.5, the sample is vortexed thoroughly andcentrifuged for 20 min at 2000 r.p.m. One ml of the upper phase is addedto 10 ml scintillation fluid, and mixed thoroughly by shaking.

[0210] The samples are then counted, 5 min per vial, in a scintillationcounter.

[0211] Lipoprotein Lipase (LPL) [Classified as EC 3.1.1.34 in accordancewith the Recommendations of the Nomenclature Committee of theInternational Union of Biochemistry and Molecular Biology (available onthe World Wide Web at the following address:http://www.chem.qmw.ac.uk/iubmb/enzyme/)]

[0212] Materials. The lipoprotein lipase employed was obtained from Dr.Gunilla Olivecrona of Ume{dot over (a)} University, Sweden. ³H-triolein(TO) from Amersham, Buckinghamshire, U.K., cat No. TRA191; 5-20 Ci/mmoldissolved in toluene. Intralipid is from Kabi-Fresenius). Apo CII wasobtained from PMSF-treated plasma from overnight fasted rats. All otherreagents are of commercial grade and obtained from various commercialsources.

[0213] Methods. The preparation of the substrate is as follows:

[0214] 25 μl of ³H-Triolein evaporated under a gentle stream of N₂ in a27° C. room in 4 ml chromacol glass followed by the addition of 1 ml 10%intralipid. This is mixed and sonicated on ice for 5 minutes (50% pulsemode). This can be kept for 1 week at 4° C.

[0215] 1. The reaction is:

[0216] 100 μl assay medium (0.3M Tris-Cl, 0.2M NaCl, 0.2 mg heparin/mland 120 mg BSA/ml (fraction V, Sigma no. 3401, pH 8.5))

[0217] 4 μl labeled intralipid

[0218] 5 μl APO CII

[0219] 86 μl MilliQ water

[0220] All mixed in a 13×100 mm glass tube and incubated for at least 5minutes.

[0221] 2. Add 5 μl LPL (of a solution containing 500 mU/mg)

[0222] Mix and incubate for 30 minutes at 25° C.

[0223] 3. The reaction is stopped with 3.25 ml of a solution containingMethanol:chloroform:heptan 10:9:7. Add 1.05 ml buffer (100 mM Boricacid, 100 mM potassium carbonate pH 10.5) and mix 10 seconds per vial.

[0224] 4. Centrifuge 20 min 12000 rpm

[0225] 5. Take 1 ml of the upper phase+10 ml scintillation fluid and mix

[0226] 6. Scintillation count for 5 minutes per vial.

[0227] Hepatic Lipase (HL) [classified as a triacylglycerol lipase, EC3.1.1.3, in accordance with the Recommendations of the NomenclatureCommittee of the International Union of Biochemistry and MolecularBiology (available on the World Wide Web at the following address:http://www.chem.qmw.ac.uk/iubmb/enzyme/)]

[0228] Materials. The hepatic lipase employed was obtained from Dr.Howard Wong of the University of California at Los Angeles (UCLA), USA,or produced and purified using the methods and a cell line provided byDr. Wong's laboratory. ³H-triolein (TO) from Amersham, Buckinghamshire,U.K., cat No. TRA191; 5-20 Ci/mmol dissolved in toluene and triolein(Sigma, Cat. No. T-1740). All other reagents are of commercial grade andobtained from various commercial sources.

[0229] Assay mix

[0230] 25 mg olive oil

[0231] 50 μl TO in a 20 ml scintillation vial and evaporate with N₂ for5 minutes.

[0232] Add 1 ml 10% gum arabic and 1.25 ml Tris-Cl pH 8.5 and 2 mlMilliQ water.

[0233] Sonicate, on ice, 16 times for 30 seconds with a 30-secondinterval between cycles.

[0234] Add 2.5 ml of 5 M NaCl+2.5 ml BSA (10% in Milli Q, pH 8)+3.25 mlMilli Q

[0235] Total volume: 12.5 ml (for 80 reactions )

[0236] If an inhibitor is going to be used, it should be incubated withthe 50 μl HL (of a 125 mU/ml solution) for 70 minutes at 25° C. A 5 μlaliquot from the incubated 50 μl is added to 150 μl assay mix+45 μlMilli Q™ water and mixed.

[0237] Incubate for exactly 30 minutes at 25° C.

[0238] Stop the reaction with Methanol:chloroform:heptane 10:9:7: 3.25ml/vial, mix; then add 1.05 ml buffer (100 mM boric acid, 100 mMpotassium carbonate pH 10.5), mix for 10 seconds per vial.

[0239] Centrifuge for 20 min/2000 rpm

[0240] Take 1 ml of the upper phase+10 ml scintillation fluid and mix byshaking by hand.

[0241] Scintillation count for 5 minutes per vial.

[0242] Results

[0243] The results of the inhibition assays for the specific compoundsof the invention described in the working examples (vide infra) aresummarized in Table 1, below. TABLE 1 HSL inhibition LPL inhibition HLinhibition Example No. (IC₅₀, μM) (% inh. at 50 μM) (% inh. at 50 μM) 10.2 5 2 6.4 2 3 0.36 0 4 9.9 20 5 0.039 7 6 0.019 0 6 7 0.46 0 8 0.03 19 1.6 7 10 0.67 5 11 >10 7 12 — 13 0.045 3 14 0.8 7 15 1.5 0 16 1.4 0 170.003 0 3 18 7.7 0 19 — 20 0.24 4 21 >10 5 22 0.03 10 23 0.04 4 4 24 >1025 >10 26 >10 27 0.09 5 28 0.07 3 29 2.6 0 30 3.8 6 31 >10 1 32 >10 0 330.17 2 34 >10 35 >10 0 36 >10 37 3.9 0 38 0.33 0 39 2.5 2 40 3.4 141 >10 42 0.04 6 43 >10 6 44 >10 45 >10 46 — 47 >10 48 0.05 49 0.10 50>10

[0244] General Methods

[0245] The compounds of the invention may be prepared by means of thecarbazate chemistry which is well known in the art, for example in M.Salman, S. Ray Indian J. Chem. Sect. B 1981, 20, 477-479; M. Marastoni,M. Bergonzoni, F. Bortolotti, R. Tomatis Arzneim. Forsch. 1997, 47,889-894; R. R. L. Hamer, R. C. Effland, J. T. Klein J. HeterocyclicChem. 1988, 25, 517; M. R. Mish, F. M. Guerra, E. M. Carreira J. Am.Chem. Soc. 1997, 119, 8379-8380; W. P. Brian J. Med. Chem. 1971, 14,1133-1134, as summarized in Schemes 1 to 3 wherein A, R¹, X¹, X² and Bare as previously defined.

[0246] wherein X¹=O, X²=O

[0247] The hydrazines II may be readily obtained by publishedmethodology, for example as described by: J. H. Biel, A. E. Drukker, T.F. Mitchell, E. P. Sprengeler, P. A. Nuhfer, A. C. Conway, A. Horita, J.Am. Chem. Soc. 1959, 81, 2805; K. Sakane, K.-i. Terayama, E. Haruki, Y.Otsuj, E. Imoto, Bull. Chem. Soc. Jpn. 1974, 47, 1297-1298; L.Landriani, D. Barlocco, G. A. Pinna, M. P. Demontis, M. M. P. Enrico, V.Anania, II Farmaco 1989, 44, 1059-1068; E. Höft, A. Rieke, Angew. Chem.1961, 73, 1297-1298.

[0248] The chloroformates and chlorodithioformates III may be readilyobtained either from commercial sources or by published methodology, forexample as described by: L. N. Owen, R. Sridhar, J. Chem. Soc (C) 1970,472475; T. Konakahara, T. Ozaki, K. Sato, B. Gold, Synthesis 1993,103-106; A.Vigroux, M. Bergon, C. Zedde, J. Med. Chem. 1995, 38,3983-3994.

[0249] Other compounds used as starting materials are either knowncompounds or compounds that can readily be prepared by methods known perse.

[0250] Preparation of the compounds of the present invention is furtherillustrated in the following examples, which are, however, not to beconstrued as limiting the invention in any way.

[0251] The structures and purities of the compounds were determined byHigh Performance Liquid Chromatography (HPLC), Nuclear MagneticResonance (NMR, Bruker 200 and 300 MHz) or Liquid Chromatography-MassSpectrometry (LC-MS). NMR chemical shifts (8) are given in parts permillion (ppm), and only selected peaks are given. Column chromatographywas carried out using the technique described by W. C. Still et al., J.Org. Chem. 1978, 43, 2923-2925, on Macherey-Nagel silica gel 60 (230-400mesh).

[0252] HPLC-Analysis:

[0253] The RP-analysis was performed using UV detection at 210 and 254nm on a YMC 120 Å 5 μl HPLC column (50×4.0 mm i.d., C-18 silica), whichwas eluted at 2.8 mL/min. The column was equilibrated with a solution oftrifluoroacetic acid (TFA) (0.01%) in water. After injection the samplewas eluted with a linear gradient of 0% to 90% acetonitrile containing0.01% TFA in the same aqueous buffer over a period of 10 min.

EXAMPLES Example 1 (3,4-Dihydro-1H-isoquinolin-2-yl)-carbamic AcidPhenyl Ester

[0254]

[0255] N,N-Diisopropylethylamine (2.41 mL) was added to a stirredsolution of 2-(2-bromomethylphenyl)ethylbromide (1.00 g) andhydrazinecarboxylic acid phenyl ester (0.94 g) in N,N-dimethylformamide(20 mL). The reaction mixture was heated at 60° C. overnight, cooleddown to room temperature and poured into water. The precipitate wasisolated by suction and recrystallised from ethyl acetate/heptaneyielding the title compound as a white solid.

[0256]¹H-NMR (300 MHz, CDCl₃); δ=3.06 (t, 2H), 3.30 (t, 2H), 4.16 (s,2H), 7.03 (m, 1H), 7.17 (m, 6H), 7.36 (m, 2H).

Example 2 (3,4-Dihydro-1H-isoquinolin-2-yl)-carbamic acid2-methoxy-phenyl Ester

[0257]

[0258] N,N-Diisopropylethylamine (2.41 mL) was added to a stirredsolution of 2-(2-bromomethylphenyl)ethylbromide (1.00 g) andhydrazinecarboxylic acid 2-methoxyphenyl ester (1.13 g) inN,N-dimethylformamide (20 mL). The reaction mixture was heated at 60° C.for 3 h, cooled down to room temperature and poured into water. Theprecipitate was isolated by suction, stripped with acetonitrile anddried at 40° C. in vacuo overnight yielding the title compound as awhite solid.

[0259]¹H-NMR (300 MHz, CDCl₃); δ=3.05 (t, 2H), 3.32 (t, 2H), 3.87 (s,3H), 4.17 (s, 2H), 6.39 (br.s, 1H), 6.94 (m, 2H), 7.03 (m, 1H), 7.14 (m,5H).

Example 3 (3,4-Dihydro-1H-isoquinolin-2-yl)-carbamic acid4-methoxy-phenyl Ester

[0260]

[0261] At 0° C., solution of 3,4dihydro-1H-isoquinolin-2-ylamine (0.50g) and N,N-diisopropylethylamine (0.58 mL) in dichloromethane (10 mL)was added dropwise to a stirred solution of 4-methoxyphenylchloroformate (0.57 mL) in dichloromethane (10 mL). After stirringovernight at room temperature the solution was extracted with water,dried over Na₂SO₄, filtered and evaporated in vacuo. The residue wasrecrystallised from ethyl acetate/heptane yielding the title compound asa white solid.

[0262]¹H-NMR (300 MHz, CDCl₃); δ=3.04 (t, 2H), 3.28 (t, 2H), 3.78 (s,3H), 4.14 (s, 2H), 6.32 (br.s, 1H), 6.86 (AB-system, 2H), 7.02 (m, 1H),7.07 (AB-system, 2H), 7.13 (m, 3H).

Example 4 4-(3,4-Dihydro-1H-isoquinolin-2-ylcarbamoyloxy)-benzoic AcidMethyl Ester

[0263]

[0264] At 0° C., solution of 3,4-dihydro-1H-isoquinolin-2-ylamine (0.50g) and N,N-diisopropylethylamine (0.58 mL) in dichloromethane (10 mL)was added dropwise to a stirred solution of 4-methoxycarbonylphenylchloroformate (0.73 g) in dichloromethane (10 mL). After stirringovernight at room temperature the solution was extracted with water,dried over Na₂SO₄, filtered and evaporated in vacuo. The off-whiteresidue was purified by flash column chromatography (SiO₂, ethylacetate/heptane 30/70 followed by ethyl acetate/heptane 20/80).Recrystallisation from ethyl acetate yielded the title compound as awhite solid.

[0265]¹H-NMR (300 MHz, CDCl₃); δ=3.05 (t, 2H), 3.32 (t, 2H), 3.92 (s,3H), 4.17 (s, 2H), 6.39 (br.s, 1H), 7.04 (m, 1H), 7.16 (m, 3H), 7.25 (d,2H), 8.06 (d, 2H).

Example 5 (3,4-Dihydro-1H-isoquinolin-2-yl)-carbamic acid 4-bromo-phenylEster

[0266]

[0267] The title compound was prepared from3,4-dihydro-1H-isoquinolin-2-ylamine hydrochloride (1.50 g) and4-bromophenyl chloroformate (1.79 g) using the procedure as described inexample 4. Purification by flash column chromatography (SiO₂, ethylacetate/heptane 30/70), followed by recrystallisation from ethyl acetateyielded the title compound as a slightly yellow solid.

[0268]¹H-NMR (300 MHz, CDCl₃); δ=3.04 (t, 2H), 3.28 (t, 2H), 4.16 (s,2H), 6.33 (br.s, 1H), 7.05 (m, 1H+d, 2H), 7.17 (m, 3H), 7.48 (d, 2H).

Example 6 (3,4-Dihydro-1H-isoquinolin-2-yl)-carbamic acid4-chloro-phenyl Ester

[0269]

[0270] The title compound was prepared from3,4-dihydro-1H-isoquinolin-2-ylamine hydrochloride (1.50 g) and4-chlorophenyl chloroformate (1.43 g) using the procedure as describedin example 4. Purification by flash column chromatography (SiO₂, ethylacetate/heptane 30/70), followed by recrystallisation from ethyl acetateyielded the title compound as a slightly yellow solid.

[0271]¹H-NMR (300 MHz, CDCl₃); δ=3.04 (t, 2H), 3.29 (t, 2H), 4.15 (s,2H), 6.35 (br.s, 1H), 7.03 (m, 1H), 7.11 (AB-system, 2H), 7.17 (m, 3H),7.33 (AB-system, 2H).

Example 7 3,4-Dihydro-1H-isoquinolin-2-yl)-carbamic Acid 4-fluoro-phenylEster

[0272]

[0273] The title compound was prepared from3,4-dihydro-1H-isoquinolin-2-ylamine (0.50 g) and 4-fluorophenylchloroformate (0.59 g) using the procedure as described in example 4.Purification by flash column chromatography (SiO₂, ethyl acetate/heptane30/70), followed by recrystallisation from ethyl acetate yielded thetitle compound as a yellow solid.

[0274]¹H-NMR (300 MHz, CDCl₃); δ=3.04 (t, 2H), 3.29 (t, 2H), 3.92 (s,3H), 4.16 (s, 2H), 6.33 (br.s, 1 H), 6.99-7.21 (m, 8H).

Example 8 (3,4-Dihydro-1H-isoquinolin-2-yl)-carbamic Acid p-tolyl Ester

[0275]

[0276] A solution of 3,4-dihydro-1H-isoquinolin-2-ylamine (0.50 g) in afew mL's of dry ether was added slowly to a stirred solution of p-tolylchloroformate (0.57 g) in dry ether (15 mL), followed by the addition ofpyridine (0.27 g) dissolved in a few mL's of dry ether. After stirringfor 5 min. at room temperature, dichloromethane was added and thesolution was extracted with water. The organic layer was dried overNa₂SO₄, filtered and evaporated in vacuo. Recrystallisation of theresidue from ethyl acetate yielded the title compound as a white solid.

[0277]¹H-NMR (300 MHz, CDCl₃); δ=2.33 (s, 3H), 3.03 (t, 2H), 3.27 (t,2H), 4.14 (s, 2H), 6.36 (br.s, 1H), 7.02 (m, 3H), 7.15 (m, 5H).

Example 9 (3,4-Dihydro-1H-isoquinolin-2-yl)-carbamic Acid o-tolyl Ester

[0278]

[0279] o-Tolyl chloroformate (0.43 g) was added to a stirred solution of3,4-dihydro-1H-isoquinolin-2-ylamine hydrochloride (0.50 g) andN,N-diisopropylethylamine (0.44 mL) in dichloromethane (10 mL). Thereaction mixture was stirred for 1 h at 0° C. followed by stirringovernight at room temperature. Extra dichloromethane was added and thesolution was extracted with water. The organic layer was dried overNa₂SO₄, filtered and evaporated in vacuo. Flash column chromatography(SiO₂, ethyl acetate/heptane 30/70), followed by recrystallisation fromethyl acetate yielded the title compound as a white solid.

[0280]¹H-NMR (300 MHz, CDCl₃); δ=2.25 (s, 3H), 3.05 (t, 2H), 3.31 (t,2H), 4.18 (s, 2H), 6.37 (br.s, 1H), 7.02-7.22 (m, 8H).

Example 10 (3,4-Dihydro-1H-isoquinolin-2-yl)-carbamic Acid m-tolyl Ester

[0281]

[0282] The title compound was prepared from3,4-dihydro-1H-isoquinolin-2-ylamine hydrochloride (0.50 g) and m-tolylchloroformate (0.43 g) using the procedure as described in example 9.

[0283]¹H-NMR (300 MHz, CDCl₃); δ=2.36 (s, 3H), 3.05 (t, 2H), 3.31 (t,2H), 4.17 (s, 2H), 6.33 (br.s, 1H), 6.93-7.25 (m, 8H).

Example 11 (3,4-Dihydro-1H-isoquinolin-2-yl)-carbamic Acid2.6-dichloro-phenyl Ester

[0284]

[0285] 2,6-Dichlorophenyl chloroformate (0.57 g) was added to a stirredsolution of 3,4-dihydro-1H-isoquinolin-2-ylamine hydrochloride (0.50 g)and N,N-diisopropylethylamine (0.44 mL) in dichloromethane (10 mL). Thereaction mixture was stirred for 1 h at 0° C. followed by stirring for 3d at room temperature. Extra dichloromethane was added and the solutionwas extracted with water. The organic layer was dried over Na₂SO₄,filtered and evaporated in vacuo. Flash column chromatography (SiO₂,ethyl acetate/heptane 30/70), followed by recrystallisation from ethylacetate yielded the title compound as a white solid.

[0286]¹H-NMR (200 MHz, CDCl₃); δ=3.04 (t, 2H), 3.33 (t, 2H), 4.20 (s,2H), 6.53 (br.s, 1H), 7.02-7.42 (m, 7H).

Example 12 (3,4-Dihydro-1H-isoquinolin-2-yl)-carbamic Acid2,4-dichloro-phenyl Ester

[0287]

[0288] 2,4-Dichlorophenyl chloroformate (0.90 g) and triethylamine (0.81g) were added respectively to a stirred suspension of3,4-dihydro-1H-isoquinolin-2-yiamine hydrochloride (0.79 g) indichloromethane (25 mL). The reaction mixture was stirred for 5 min at0° C. followed by stirring at room temperature. After 2.5 h an extraequivalent of 2,4-dichlorophenyl chloroformate was added, followed by athird equivalent after another 2 h. After stirring overnight thesolution was extracted with water. The organic layer was dried overNa₂SO₄, filtered and evaporated in vacuo. Flash column chromatography(SiO₂, ethyl acetate/heptane 30/70), followed by recrystallisation fromethyl acetate yielded the title compound as a white solid.

[0289]¹H-NMR (300 MHz, CDCl₃); δ=3.06 (t, 2H), 3.31 (t, 2H), 4.17 (s,2H), 6.47 (br.s, 1H), 7.03 (m, 1H), 7.10-7.28 (m, 5H), 7.43 (d, 1H).

Example 13 (3,4-Dihydro-1H-isoquinolin-2-yl)-carbamic Acid4-trifluoromethyl-phenyl Ester

[0290]

[0291] The title compound was prepared from3,4-dihydro-1H-isoquinolin-2-ylamine (0.50 g) and4-trifluoromethylphenyl chloroformate (0.76 g) using the procedure asdescribed in example 8.

[0292]¹H-NMR (300 MHz, CDCl₃); δ=3.04 (t, 2H), 3.30 (t, 2H), 4.15 (s,2H), 7.03 (m, 1H), 7.15 (m, 3H), 7.39 (d, 2H), 7.63 (d, 2H).

Example 14 (3,4-Dihydro-1H-isoquinolin-2-yl)-carbamic Acid2-bromo-phenyl Ester

[0293]

[0294] 3,4-Dihydro-1H-isoquinolin-2-ylamine hydrochloride (0.79 g) andtriethylamine (0.81 g) were added respectively to a stirred solution of2-bromophenyl chloroformate (0.94 g) in dichloromethane (25 mL). Afterstirring for 0.5 h water was added and both layers were separated. Theorganic layer was dried over Na₂SO₄, filtered and evaporated in vacuoyielding a white solid. Flash column chromatography (SiO₂, ethylacetate), followed by recrystallisation from ethyl acetate andchloroform yielded the title compound as a white solid.

[0295]¹H-NMR (300 MHz, CDCl₃); δ=3.06 (t, 2H), 3.32 (t, 2H), 4.18 (s,2H), 6.50 (br.s, 1H), 7.00-7.35 (m, 7H), 7.58 (dd, 1H).

Example 15 (3,4-Dihydro-1H-isoquinolin-2-yl)-carbamic Acid2-fluoro-phenyl Ester

[0296]

[0297] The title compound was prepared from3,4-dihydro-1H-isoquinolin-2-ylamine hydrochloride (0.79 g) and2-fluorophenyl chloroformate (0.70 g) using the procedure as describedin example 14.

[0298]¹H-NMR (300 MHz, CDCl₃); δ=3.03 (t, 2H), 3.29 (t, 2H), 4.17 (s,2H), 6.48 (br.s, 1H), 7.03 (m, 1H), 7.08-7.27 (m, 7H).

Example 16 (6,7-Dimethoxy-3.4-dihydro-1H-isoquinolin-2-yl)-carbamic Acid4-chloro-phenyl Ester

[0299]

[0300] Triethylamine (0.17 mL) was added to a stirred suspension of6,7-dimethoxy-3,4-dihydro-1H-isoquinolin-2-ylamine hydrochloride (0.30g) and 4-chlorophenyl chloroformate (0.23 g) in dichloromethane (10 mL).After stirring for 3 d at room temperature additional dichloromethanewas added and the solution was extracted with water. The water layer wasextracted with dichloromethane and the combined organic layers weredried over Na₂SO₄, filtered and evaporated in vacuo. Recrystallisationfrom ethyl acetate yielded the title compound as a white solid.

[0301]¹H-NMR (300 MHz, CDCl₃); δ=2.96 (t, 2H), 3.27 (t, 2H), 3.85 (s,3H), 3.86 (s, 3H), 4.07 (s, 2H), 6.38 (br.s, 1H), 6.51 (s, 1H), 6.62 (s,1H), 7.12 (AB-system, 2H), 7.33 (AB-system, 2H).

Example 17 (3,4-Dihydro-1H-isoquinolin-2-yl)-carbamic Acid4-tert-butoxycarbonylamino-phenyl Ester

[0302]

[0303] (4-Hydroxyphenyl)-carbamic acid tert-butyl ester (355 mg) andpyridine (284 mg) were dissolved in dichloromethane (20 mL). A solutionof triphosgene (168 mg) in dichloromethane (10 mL) was added. Afterstirring for 1 h at 0° C., a solution of3,4-dihydro-1H-isoquinolin-2-ylamine hydrochloride (315 mg) andtriethylamine (172 mg) in dichloromethane (10 mL) was added. The coolingbath was removed and the solution was stirred overnight at roomtemperature. A small amount of triethylamine was added and stirring wascontinued for 1 h. The solution was extracted with water (2×), driedover Na₂SO₄, filtered and evaporated in vacuo. Purification by flashcolumn chromatography (SiO₂, ethyl acetate/heptane 30/70) followed byrecrystallisation from ethyl acetate/heptane yielded the title compoundas a white solid.

[0304]¹H-NMR (300 MHz, CDCl₃); δ=1.52 (s, 9H), 3.04 (t, 2H), 3.28 (t,2H), 4.14 (s, 2H), 6.32 (br.s, 1H), 6.47 (br.s, 1H), 7.00-7.18 (m, 6H),7.33 (d, 2H).

[0305] LC-MS (electrospray) m/z: 384 (M+H)⁺, 767 (2M+H)⁺.

Example 18 (3,4-Dihydro-1H-isoquinolin-2-yl)-carbamic Acid4-amino-phenyl Ester Hydrochloride

[0306]

[0307] A solution of (3,4-dihydro-1H-isoquinolin-2-yl)-carbamic acid4-tert-butoxycarbonylamino-phenyl ester (32 mg) in a mixture oftetrahydrofuran/formic acid/1N hydrochloric acid (5 mL) was stirredovernight at room temperature. The solvent was evaporated under reducedpressure and the residue was recrystallised from ethanol yielding thetitle compound as a white solid.

[0308]¹H-NMR (300 MHz, MeOH-d₄); δ=3.05 (t, 2H), 3.19 (t, 2H), 4.07 (s,2H), 7.06 (m, 1H), 7.12 (m, 3H), 7.33 (AB-system, 2H), 7.42 (AB-system,2H).

[0309] LC-MS (electrospray) m/z: 284.0 (M+H)⁺, 589.3 (2M+Na)⁺.

Example 19 4-(3,4-Dihydro-1H-isoquinolin-2-ylcarbamoyloxy)-benzoic AcidBenzyl Ester

[0310]

[0311] Pyridine (237 mg) was added carefully to a stirred solution of4-chlorocarbonyloxy-benzoic acid benzyl ester (1.31 g) and3,4-dihydro-1H-isoquinolin-2-ylamine hydrochloride (554 mg) indichloromethane (10 mL). After stirring for 24 h at room temperatureextra dichloromethane was added and the solution was extracted withwater. The organic layer was dried over Na₂SO₄, filtered and evaporatedin vacuo. The residue was purified by flash column chromatography (SiO₂,ethyl acetate/heptane 20/80) yielding the title compound as a whitesolid.

[0312]¹H-NMR (300 MHz, CDCl₃); δ=3.06 (t, 2H), 3.29 (t, 2H), 4.16 (s,2H), 5.35 (s, 2H), 6.45 (br.s, 1H), 7.03 (m, 1H), 7.16 (m, 3H), 7.23 (d,2H), 7.33-7.47 (m, 5H), 8.08 (d, 2H),

[0313] LC-MS (electrospray) m/z: 403 (M+H)⁺, 827 (2M+Na)⁺.

Example 20 (3,4-Dihydro-1H-isoquinolin-2-yl)-carbamic Acid4-propyl-phenyl Ester

[0314]

[0315] At 0° C., 4-n-propylphenyl chloroformate (0.79 g) andtriethylamine (0.81 g) were added respectively to a stirred suspensionof 3,4-dihydro-1H-isoquinolin-2-ylamine hydrochloride (0.79 g) indichloromethane (25 mL). The cooling bath was removed and the reactionmixture was stirred overnight at room temperature. Another equivalent of4-n-propylphenyl chloroformate was added and stirring was continued foranother 2.5 h. The solution was extracted with water, dried over Na₂SO₄,filtered and evaporated in vacuo, yielding a slightly yellow powder. Theresidue was purified by flash column chromatography (SiO₂, ethylacetate/heptane 30/70). Recrystallisation from ethyl acetate yielded thetitle compound as a white solid.

[0316]¹H-NMR (300 MHz, CDCl₃); δ=0.93 (t, 3H), 1.63 (m, 2H), 2.56 (t,2H), 3.03 (t, 2H), 3.28 (t, 2H), 4.16 (s, 2H), 6.34 (br.s, 1H), 7.02 (m,3H), 7.15 (m, 5H).

Example 21 (3,4-Dihydro-1H-isoquinolin-2-yl)-methyl-carbamic Acid4-chloro-phenyl Ester

[0317]

[0318] Triethylamine (0.42 mL) was added to a stirred suspension of(3,4-dihydro-1H-isoquinolin-2-yl)-methylamine hydrochloride (200 mg) indichloromethane (10 mL), followed by a dropwise addition of4-chlorophenyl chloroformate (211 mg). After stirring overnight at roomtemperature, extra dichloromethane was added and the solution wasextracted with water. The organic layer was dried over Na₂SO₄, filteredand evaporated in vacuo. The residue was purified by flash columnchromatography (SiO₂, ethyl acetate/heptane 10/90) yielding the titlecompound as a white solid.

[0319]¹H-NMR (300 MHz, CDCl₃); δ=2.8-3.5 (br.m, 7H), 4.24.4 (br.d, 2H),7.00-7.18 (m, 6H), 7.30 (d, 2H).

Example 22 (1-Methyl-3,4-dihydro-1H-isoquinolin-2-yl)-carbamic Acid4chloro-phenyl Ester

[0320]

[0321] Triethylamine (0.84 mL) was added dropwise to a stirredsuspension of 1-methyl-3,4-dihydro-1H-isoquinolin-2-ylaminehydrochloride (0.60 g) and 4-chlorophenyl chloroformate (0.63 g) indichloromethane (20 mL). After stirring for 2 h at room temperature,extra dichloromethane was added and the solution was extracted twicewith water. The organic layer was dried over Na₂SO₄, filtered andevaporated in vacuo. The residue was recrystallised twice from ethylacetate yielding the title compound as a white solid.

[0322]¹H-NMR (300 MHz, CDCl₃); δ=1.55 (d, 3H), 2.83-3.49 (m, 4H), 4.15(br.s, 1H), 6.30 (br.s 1 H), 7.07-7.22 (m, 6H), 7.32 (d, 2H).

Example 23 (3-Methyl-3,4-dihydro-1H-isoquinolin-2-yl)-carbamic Acid4-chloro-phenyl Ester

[0323]

[0324] At 0° C., N,N-diisopropylethylamine (781 mg) was added to astirred solution of 3-methyl-3,4-dihydro-1H-isoquinolin-2-ylaminehydrochloride (600 mg) and 4-chlorophenyl chloroformate (577 mg) indichloromethane (10 mL). The cooling bath was removed and stirring wascontinued for 3 h at room temperature. The reaction mixture wasextracted with water (2×), dried over Na₂SO₄, filtered and evaporated invacuo. Purification of the residue by flash column chromatographyyielded the title compound as a white solid.

[0325]¹H-NMR (300 MHz, CDCl₃); δ=1.31 (d, 3H), 2.80 (dd, 1H), 2.88 (dd,1H), 3.32 (br.s, 1H), 4.17 (d, 1H), 4.31 (br.d, 1 H), 6.21 (br.s, 1H),7.06 (m, 4H), 7.18 (m, 2H), 7.30 (d, 2H).

[0326] LC-MS (electrospray) m/z: 317 (M+H)⁺.

Examples 24-47

[0327]

[0328] The 4-(4-formyl-3,5-dimethoxyphenoxy)butyryl aminomethylpolystyrene resin (loading 0.62 mMol/g, 3.0 g) was swollen indichloromethane for 0.5 min. The solvent was removed and a solution of3,4-dihydro-1H-isoquinolin-2-ylamine hydrochloride (3.43 g) in a mixtureof N-methylpyrolidinone (30 mL) and water (1.5 mL) was added, followedby sodium cyanoborohydride (1.73 g) in N-methylpyrolidinone (10 mL) andacetic acid (4 mL). The reaction mixture was shaken overnight at roomtemperature. The solvent was removed and the resin was washed withN-methylpyrolidinone (3×30 mL), dichloromethane (3×30 mL) and methanol(3×30 mL), followed by drying overnight at 40° C. in a vacuum oven. Theresin (2.63 g) was swollen in dichloromethane for 0.5 min. Triphosgene(1.45 g) dissolved in dichloromethane (30 mL), andN,N-diisopropylethylamine (2.11 g) were added respectively. The reactionmixture was shaken for 4 h at room temperature, washed withdichloromethane (3×) and diethyl ether (3×) and dried overnight at 40°C. in a vacuum oven. A suspension of the resin was made in a 1:1 mixtureof 1,2-dichloropropane and N-methylpyrolidinone (50 mL), which wasdivided equally into 24 reaction vessels. The solvent was removed andthe resin was washed with N,N-dimethylformamide.1,4-Diazabicyclo[2.2.2]octane (10 equivalents for each reaction vessel)dissolved in N,N-dimethylformamide (25 mL) was divided equally into the24 reaction vessels. The reaction mixtures were shaken for 0.5 h at roomtemperature. The 24 different phenols were each dissolved inN,N-dimethylformamide (1 mL) and added to the 24 reaction vessels. Aftershaking overnight the solvent was removed and the resin was washed withN,N-dimethylformamide (5×) and dichloromethane (10×) respectively. Thevessels were each treated with dichloromethane (1.5 mL) andtrifluoroacetic acid (0.5 mL) for 1 h. The filtrates were collected andevaporated in vacuo yielding 24 compounds as solids or as highly viscousoils. The compounds were analysed by LC-MS in which all 24 compoundsshowed the expected molecular ion. EXAMPLE PRODUCT Formula MW LC-MSEXAMPLE 24

C₁₉H₂₀N₂O₃ 324.38 325 EXAMPLE 25

C₁₈H₉N₃O₃ 325.37 326 EXAMPLE 26

C₂₄H₂₂N₂O₃ 386.45 388 EXAMPLE 27

C₂₃H₂₂N₂O₂ 358.44 359 EXAMPLE 28

C₁₈H₂₀N₂O₂ 296.37 297 EXAMPLE 29

C₂₃H₂₂N₂O₃ 374.44 376 EXAMPLE 30

C₂₀H₂₄N₂O₃ 340.42 341 EXAMPLE 31

C₂₁H₂₆N₂O₂ 338.45 340 EXAMPLE 32

C₂₀H₂₄N₂O₂ 324.42 325 EXAMPLE 33

C₂₀H₂₄N₂O₂ 324.42 325 EXAMPLE 34

C₂₀H₂₅N₃O₂ 339.44 340 EXAMPLE 35

C₁₉H₂₀N₂O₄ 340.38 341 EXAMPLE 36

C₂₂H₂₀N₂O₂ 344.41 345 EXAMPLE 37

C₂₁H₂₆N₂O₃ 354.45 355 EXAMPLE 38

C₂₂H₂₀N₂O₃ 360.41 361 EXAMPLE 39

C₁₉H₂₂N₂O₃ 326.39 327 EXAMPLE 40

C₁₇H₁₅F₃N₂O₃ 352.31 353 EXAMPLE 41

C₂₃H₂₀N₂O₃ 372.42 373 EXAMPLE 42

C₁₉H₂₀N₂O₄ 340.38 341 EXAMPLE 43

C₁₉H₂₀N₂O₄ 350.46 351 EXAMPLE 44

C₂₀H₂₁ClN₂O₃ 372.85 373 EXAMPLE 45

C₁₈H₁₈N₂O₃ 310.35 311 EXAMPLE 46

C₁₆H₁₄Cl₂N₂O₂ 337.20 337 EXAMPLE 47

C₁₇H₁₄ClF₃N₂O₂ 370.76 371

Example 48 Piperidin-1-yl-carbamic Acid 4-chloro-phenyl Ester

[0329]

[0330] 4-Chlorophenyl chloroformate (2.84 g) and triethylamine (2.09 mL)were added respectively to a stirred solution of N-aminopiperidine (1.50g) in dichloromethane (30 mL). After stirring for 3 d at roomtemperature the solution was extracted with water. The organic layer wasdried over Na₂SO₄, filtered and evaporated in vacuo. Purification byflash column chromatography (ethyl acetate/heptane 20/80) yielded thetitle compound as a white solid.

[0331]¹H-NMR (300 MHz, CDCl₃); δ=1.42 (m, 2H), 1.72 (m, 4H), 2.83 (m,4H), 5.96 (br.s, 1H), 7.08 (d, 2H), 7.28 (d, 2H).

[0332] LC-MS (electrospray) m/z: 255 (M+H)⁺.

Example 49 Morpholin-4-yl-carbamic Acid 4-chloro-phenyl Ester

[0333]

[0334] 4-Chlorophenyl chloroformate (1.15 g) was added slowly to astirred solution of N-aminomorpholine (613 mg) and pyridine (475 mg) indichloromethane. After stirring overnight dichloromethane was added andthe solution was extracted with water, dried over Na₂SO₄, filtered andevaporated in vacuo. Recrystallisation of the residue from ethylacetate/heptane yielded the title compound as a white solid.

[0335]¹H-NMR (300 MHz, CDCl₃); δ=2.92 (m, 4H), 3.81 (m, 4H), 6.14 (br.s,1H), 7.08 (AB-system, 2H), 7.32 (AB-system, 2H).

[0336] LC-MS (electrospray) m/z: 257 (M+H)⁺.

Example 50 (3,4-Dihydro-1H-isoquinolin-2-yl)-dithiocarbamic Acid PhenylEster

[0337]

[0338] Phenyl chlorodithioformate (0.53 g) and 4-dimethylaminopyridine(0.66 g) were added to a stirred solution of3,4-dihydro-1H-isoquinolin-2-ylamine hydrochloride (0.50 g) inN,N-dimethylformamide (10 mL). The reaction mixture was stirred for 5 dat room temperature. Flash column chromatography (SiO₂, ethylacetate/heptane 30/70), followed by recrystallisation from ethylacetate/heptane yielded the title compound as a white solid.

[0339]¹H-NMR (300 MHz, DMSO-d₆); δ=2.82-3.22 (m, 3H), 3.32 (br.s, 1H),4.03 (br. AB-system, 1H), 4.19 (br. AB-system, 1H), 7.16 (m, 4H), 7.43(s, 5H), 11.27 (s, 1H).

[0340] LC-MS (electrospray) m/z: 301 (M+H)⁺.

1. A compound of the general formula I

wherein A is a nitrogen containing ring system attached through thenitrogen atom, which nitrogen containing ring system is optionallysubstituted with one or more substituents independently selected fromhalogen, hydroxy, amino, oxy, cyano, nitro, C₁₋₆-alkyl and C₁₋₆-alkoxy;wherein each of the C₁₋₆-alkyl or C₁₋₆-alkoxy may optionally besubstituted with one or more substituents independently selected fromhydroxy, halogen, amino, cyano and nitro; R¹ is hydrogen or C₁₋₆-alkyl;X¹ is —S— or —O—; X² is —S— or —O—; B is aryl optionally substitutedwith one or more substituents independently selected from halogen,hydroxy, amino, cyano, nitro, C₁₋₆-alkyl, C₁₋₆-alkoxy,C₁₋₆-alkylcarbonyl, C₁₋₆-alkoxycarbonyl, C₃₋₁₀-cycloalkyl, aryl,aryl-C₁₋₆-alkyl, aryl-C₁₋₆-alkoxy, aryl-C₁₋₆-alkoxycarbonyl,arylcarbonyl, —NR²⁰—C(═O)—C₁₋₆-alkyl, —NR²⁰—C(═O)—C₁₋₆-alkoxy,—NR²⁰—C₁₋₆-alkyl and —C₁₋₆-alkyl-NR²⁰—C₁₋₆-alkyl; wherein each of theC₁₋₆-alkyl, C₁₋₆-alkoxy, C₃₋₁₀-cycloalkyl or aryl may optionally besubstituted with one or more substituents independently selected fromhydroxy, halogen, amino, cyano and nitro; R²⁰ is hydrogen or C₁₋₆-alkyl;or B is

wherein Y¹ is —C(R²¹)═ or —N═; Y² is —C(R²²)═ or —N═; Y³ is —C(R²³)═ or—N═; Y⁴ is —C(R²⁴)═ or —N═; wherein R²¹, R²², R²³ and R²⁴ independentlyare hydrogen, halogen, hydroxy, amino, cyano, nitro, C₁₋₆-alkyl,C₁₋₆-alkoxy, C₁₋₆-alkylcarbonyl, C₁₋₆-alkoxycarbonyl, C₃₋₁₀-cycloalkyl,aryl, aryl-C₁₋₆-alkyl, aryl-C₁₋₆-alkoxy, aryl-C₁₋₆-alkoxycarbonyl,arylcarbonyl, —NR²⁵—C(═O)—C₁₋₆-alkyl, —NR²⁵—C(═O)—C₁₋₆-alkoxy,—NR²⁵—C₁₋₆-alkyl or —C₁₋₆-alkyl-NR²⁵—C₁₋₆-alkyl; wherein each of theC₁₋₆-alkyl, C₁₋₆-alkoxy, C₃₋₁₀-cycloalkyl or aryl may optionally besubstituted with one or more substituents independently selected fromhydroxy, halogen, amino, cyano and nitro; R²⁵ is hydrogen or C₁₋₆-alkyl;or a pharmaceutically acceptable salt thereof.
 2. The compound accordingto claim 1, wherein A is

wherein R⁷, R⁸ and R⁹ independently are hydrogen or C₁₋₆-alkyl; R¹⁰,R¹¹, R¹² and R¹³ independently are hydrogen, halogen, C₁₋₆-alkyl orC₁₋₆-alkoxy.
 3. The compound according to claim 1, wherein A is

wherein R¹⁴, R¹⁵, R¹⁶, R¹⁷ and R¹⁸ independently are hydrogen, halogen,C₁₋₆-alkyl or C₁₋₆-alkoxy.
 4. The compound according to any one of thepreceding claims, wherein X¹ is —O—, and X² is —O—.
 5. The compoundaccording to any one of the preceding claims, wherein R¹ is hydrogen ormethyl.
 6. The compound according to any one of the preceding claims,wherein B is

wherein R², R³, R⁴, R⁵ and R⁶ independently of each other are hydrogen,halogen, hydroxy, amino, cyano, nitro, C₁₋₆-alkyl, C₁₋₆-alkoxy,C₁₋₆-alkylcarbonyl, C₁₋₆-alkoxycarbonyl, C₃₋₁₀-cycloalkyl, aryl,aryl-C₁₋₆-alkyl, aryl-C₁₋₆-alkoxy, aryl-C₁₋₆-alkoxycarbonyl,arylcarbonyl, —NR²⁰—C(═O)—C₁₋₆-alkyl, —NR²⁰—C(═O)—C₁₋₆-alkoxy,—NR²⁰—C₁₋₆-alkyl or —C₁₋₆-alkyl-NR²⁰—C₁₋₆-alkyl; wherein each of theC₁₋₆-alkyl, C₁₋₆-alkoxy, C₃₋₁₀-cycloalkyl or aryl may optionally besubstituted with one or more substituents independently selected fromhydroxy, halogen, amino, cyano and nitro; R²⁰ is hydrogen or C₁₋₆-alkyl.7. The compound according to any one of the preceding claims, wherein Bis


8. The compound according to claim 7, wherein B is

wherein R¹⁹ is hydrogen, C₁₋₆-alkyl, C₁₋₆-alkylcarbonyl,C₁₋₆-alkoxycarbonyl, aryl-C₁₋₆-alkyl, aryl-C₁₋₆-alkoxycarbonyl orarylcarbonyl.
 9. The compound according to any one of the precedingclaims, selected from

and pharmaceutically acceptable salts thereof.
 10. A pharmaceuticalcomposition comprising, as an active ingredient, a compound as definedin any one of the preceding claims, or a pharmaceutically acceptablesalt thereof, together with a pharmaceutically acceptable carrier ordiluent.
 11. The composition according to claim 10 in unit dosage form,comprising from about 0.05 mg to about 2000 mg, preferably from about0.1 mg to about 500 mg of the compound according to any one of claims1-9 or pharmaceutically acceptable salt thereof.
 12. A pharmaceuticalcomposition for specifically inhibiting the lipolytic activity of HSL,said composition comprising, as an active ingredient, a compoundaccording to any one of claims 1-9, or a pharmaceutically acceptablesalt thereof, together with a pharmaceutically acceptable carrier ordiluent.
 13. A pharmaceutical composition according to any one of claims10-12 for oral, nasal, transdermal, pulmonary or parenteraladministration.
 14. Use of a compound that specifically inhibits thelipolytic activity of HSL, or a pharmaceutically acceptable saltthereof, for the preparation of a medicament for the treatment of adisorder where a decreased level of plasma FFA is desired.
 15. The useaccording to claim 14, wherein said disorder where a decreased level ofplasma FFA is desired is diabetes type 2, insulin resistance, impairedglucose tolerance, hyperglycemia, dyslipidemia, or abnormalities oflipoprotein metabolism.
 16. The use according to claim 14 or 15, whereinsaid compound that specifically inhibits lipolytic activity of HSL is acompound according to any one of claims 1-9 or a pharmaceuticallyacceptable salt thereof.
 17. Use of a compound according to any one ofclaims 1-9, or a pharmaceutically acceptable salt thereof, for thepreparation of a medicament.
 18. A method of treating a disorder in amammal where a decreased level of plasma FFA is desired, said methodcomprising administering to said mammal an effective amount of acompound that specifically inhibits the lipolytic activity of HSL, or apharmaceutically acceptable salt thereof.
 19. The method according toclaim 18, wherein said disorder where a decreased level of plasma FFA isdesired is diabetes type 2, insulin resistance, impaired glucosetolerance, hyperglycemia, dyslipidemia, or abnormalities of lipoproteinmetabolism.
 20. The method according to claim 18 or 19, wherein saidcompound that specifically inhibits lipolytic activity of HSL is acompound according to any one of claims 1-9 or a pharmaceuticallyacceptable salt thereof.
 21. The method according to any one of claims18-20, wherein said administration is carried out by the oral, nasal,transdermal, pulmonary or parenteral route.
 22. A method for identifyingcompounds for the treatment of disorders where a decreased level ofplasma FFA is desired, characterised by screening out compounds thatspecifically inhibit HSL.